Roughly cylindrical sample containers having multiple reservoirs therein and being adapted for acoustic ejections

ABSTRACT

Provided herein is generally tubular container, preferably including a plurality of reservoirs defined therein. The container can be adapted for acoustic ejection of a fluid disposed within at least one of the reservoirs of the plurality of reservoirs. Alternatively, the container can be adapted for extraction of a fluid disposed within at least one of the reservoirs of the plurality of reservoirs using a non-acoustic liquid handling method.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation under 35 U.S.C. § 120 of U.S. patentapplication Ser. No. 14/595,063, filed Jan. 12, 2015 and entitled“Roughly Cylindrical Sample Containers Having Multiple ReservoirsTherein and Being Adapted for Acoustic Ejections,” which claims thebenefit of U.S. Provisional Patent Application No. 61/927,878, filed onJan. 15, 2014, the entire contents of each of which are incorporated byreference herein.

BACKGROUND OF THE INVENTION

The present invention is directed to sample handling. More particularly,certain embodiments of the present invention provide sample containersadapted for acoustic ejections and analyses and methods thereof as wellas containing multiple reservoirs. Merely by way of example, theinvention has been applied to a biological or chemical sample containerwherein multiple fluid samples, which preferably but not necessarily arerelated to one another, may be stored such as different concentrationsof the same chemical, different fractions of a patient blood sample(e.g., plasma, buffy coat, erythrocytes) in a manner compatible withboth acoustic ejection and sample handling equipment for single samplestorage and retrieval. But it would be recognized that the invention hasa much broader range of applicability and could be applied to anycollection of samples where the retrieval of a group of samples wouldspeed throughput by reducing the number of container storage andretrieval operations, increase the density of sample storage or allowfor a larger number of aliquots of the same sample to be preserved.

It is often desired to take a chemical or biological sample (e.g., ahuman blood sample) contained in an individual container and to transferit to one or more well plates or other objects appropriate for carryingout reactions and assays such as in high-throughput screening for drugdiscovery or in clinical diagnostics in automated clinical chemistryanalyzers. An important feature for the handling of samples includes theability to transfer small volumes from the container to enable varioustypes of diagnostics that can benefit from consistent deliveries ofsmall-volume samples and to be able to repeatedly extract sample fromthe same container.

Acoustic ejection has been known for a number of years as a way ofperforming transfers of samples from containers, including microplatesand microtubes. For example, in a typical setup for acoustic ejection, apiezoelectric transducer is driven by a waveform chosen by a controllerand in response generates acoustic energy. The acoustic energy often isfocused by an acoustic lens, and coupled to a reservoir or containercontaining fluid through an acoustic coupling medium (e.g., water). Ifthe focused energy has a focal point inside a fluid in the container andclose to a free surface of that fluid, a droplet may be ejected. Dropletsize and velocity can be controlled by the chosen waveform as mentionedabove.

In some embodiments, the transducer is movable in one or more directions(e.g., in the “z direction”) that is roughly perpendicular to the freesurface of the fluid. The movement can take place under the control ofthe controller. Some acoustic instruments for high-throughput use relyon an active control of the transducer position relative to thecontainer and address the multiplicity of reservoirs in microplates orto an individual tube or to a tube in a rack of tubes. Often, theadjustment of the transducer position involves sending a motion commandto a motion controller which then initiates movement in one or moredirections (e.g., along one or more axes). For example, motion in thehorizontal plane (e.g., in the “x direction” and/or in the “ydirection”) aligns the transducer with the selected reservoir, andmotion in the vertical direction (e.g., in the “z direction”) is usedboth to audit the reservoir and to focus for droplet transfer. Inanother example, positioning of the transducer to achieve the properfocus for droplet ejections can be responsive to data collected from anacoustic audit. Additionally, U.S. Pat. Nos. 6,938,995 and 7,900,505 areincorporated by reference herein for all purposes. When the motion iscomplete, the controller can notify the system that the transducer andthe selected reservoir are now in the proper position for the next stepin the process. This may be further measurement of the fluid in thereservoir and/or acoustic ejection of droplets. When completed, thefirst reservoir is removed, and the acoustic coupling with a secondreservoir may take place. Coupling fluid may remain attached to thefirst reservoir and would typically be at the surface facing thetransducer.

Containers may include one or more fluid reservoirs. For example, acontainer may include one reservoir such as individual tubes, or mayinclude a rack of separable tubes, or may include a microplate havingnon-separable wells. Individual tubes with one reservoir and microplateswith multi-reservoirs are common, and the infrastructure for supportingthe storage, retrieval and use of such tubes and plates is also common.

As is known in the art, an advantage of a single tube is that the sampletherein can be stored and retrieved independently of other samples, andan advantage of a microplate is that it can store and retrieve a largenumber (96, 385, 1536, 3456) of samples which can be small in volume(e.g., under 1 μL for the highest density microplates).

SUMMARY OF THE INVENTION

The present invention is directed to sample handling. More particularly,certain embodiments of the present invention provide sample containersadapted for acoustic ejections and analyses and methods thereof as wellas containing multiple reservoirs. Merely by way of example, theinvention has been applied to a biological or chemical sample containerwherein multiple fluid samples, which preferably but not necessarily arerelated to one another, may be stored such as different concentrationsof the same chemical, different fractions of a patient blood sample(e.g., plasma, buffy coat, erythrocytes) in a manner compatible withboth acoustic ejection and sample handling equipment for single samplestorage and retrieval. But it would be recognized that the invention hasa much broader range of applicability and could be applied to anycollection of samples where the retrieval of a group of samples wouldspeed throughput by reducing the number of container storage andretrieval operations, increase the density of sample storage or allowfor a larger number of aliquots of the same sample to be preserved.

For example, there is a need for individual sample containers with theadvantages of being both compatible with acoustic ejection systems thatare amenable to miniaturization and can provide multiple reservoirs froma single retrieval. In particular, it would be desirable for thecontainer to hold related materials that would be likely to be retrievedand used for a specific function, such as an assay of different samplesfrom the same patient (e.g., different fractionations of blood ortime-course samples) or various concentrations of the same compound forconstruction of dose response curves.

Accordingly, some embodiments of the present invention provide anindividual, generally cylindrical container that is configured tocontain multiple different samples. Optionally, the container may becombined with other containers, e.g., stored in association with othercontainers, e.g., stacked with other containers, for retrieval as agroup. In one embodiment, one or more of these combinable cylindricalcontainers with multiple reservoirs could also be combined with at leastone cylindrical container having only one reservoir.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A-1B are simplified diagrams showing a single retrievable samplecontainer of roughly cylindrical shape having more than one reservoirwhere the samples are isolated from each other, optionally including anacoustically distinguishable difference in thickness in the bottoms ofreservoirs in the container, according to some embodiments of thepresent invention.

FIG. 2 is a simplified diagram showing a retrievable sample container ofroughly cylindrical shape having more than one reservoir where thesamples are isolated from each other, in conjunction with an acousticejection system, according to some embodiments of the present invention.

FIGS. 3A-3C illustrate exemplary configurations for storing samplecontainers with multiple reservoirs therein either singly or incombination with other containers, e.g., stacked with other containers,according to some embodiments of the present invention.

FIG. 4A shows illustrates an exemplary rack for storing combinations,e.g., stacks of roughly cylindrical containers with multiple reservoirstherein for storage and retrieval, and FIG. 4B illustrates an exemplaryrack for individually holding roughly cylindrical containers withmultiple reservoirs therein for analysis or acoustic ejection, accordingto some embodiments of the present invention.

FIG. 5A-5B are simplified diagrams showing exemplary collections ofmultiple sample containers surface with their corresponding seals (orlids) stacked as a linked assembly with roughly cylindrical exterior forstorage or retrieval with different concentrations in each well,according to some embodiments of the present invention.

DETAILED DESCRIPTION

The present invention is directed to sample handling. More particularly,certain embodiments of the present invention provide sample containersadapted for acoustic ejections and analyses and methods thereof as wellas containing multiple reservoirs. Merely by way of example, theinvention has been applied to a biological or chemical sample containerwherein multiple fluid samples, which preferably but not necessarily arerelated to one another, may be stored such as different concentrationsof the same chemical, different fractions of a patient blood sample(e.g., plasma, buffy coat, erythrocytes) in a manner compatible withboth acoustic ejection and sample handling equipment for single samplestorage and retrieval. But it would be recognized that the invention hasa much broader range of applicability and could be applied to anycollection of samples where the retrieval of a group of samples wouldspeed throughput by reducing the number of container storage andretrieval operations, increase the density of sample storage or allowfor a larger number of aliquots of the same sample to be preserved.

With respect to various embodiments of the present invention, it is tobe understood that this invention is not limited to specific solvents,materials, and/or device structures, as such may vary. It is also to beunderstood that the terminology used herein is for the purpose ofdescribing particular embodiments only, and is not intended to belimiting.

According to some embodiments, the singular forms “a,” “an,” and “the”include both singular and plural referents unless the context clearlydictates otherwise. For example, reference to “a fluid” includes aplurality of fluids as well as a single fluid. In another example,reference to “a temperature” includes a plurality of temperatures aswell as a single temperature.

According to certain embodiments, where a range of values is provided,it is intended that each intervening value between the upper limit andthe lower limit of that range and any other stated or intervening valuein that stated range is encompassed within the disclosure. For example,if a range of 1 μm to 8 μm is stated, it is intended that at least 2 μm,3 μm, 4 μm, 5 μm, 6 μm, and 7 μm are also disclosed, as well as therange of values that are greater than or equal to 1 μm and less than orequal to 8 μm.

According to some embodiments, reference is sometimes made to“horizontal” or “vertical” in terms of acoustic ejection configurationwhere a fluid is in a sample container and has a free surface which isapproximately horizontal (e.g., approximately perpendicular to thedirection of the earth's gravity).

As discussed above, there is a need for sample containers that cansimplify the full life cycle of processing biological samples (e.g.,collecting, transferring, preserving, and/or analyzing biologicalsamples) by using acoustic ejection and/or acoustic analysis, and thatinclude multiple reservoirs that respectively may be configured to storemultiple fluids. In some embodiments, the multiple reservoirsrespectively are configured to store multiple different concentrationsof a particular fluid.

For example, because acoustics allows for smaller reservoirs as well asthe volumes extracted from them, having multiple reservoirs definedwithin a generally cylindrical container of conventional size is bothpossible and useful. For example, as provided herein, a containerconfigured for use within a conventional microplate footprint, e.g., amicroplate having 96, 385, 1536, or 3456 wells, such as a 1536 lowdead-volume (LDV) microplate available from Labcyte Inc. (Sunnyvale,Calif.) or any other suitable microplate or rack, and configured forstorage and retrieval using conventional robotic systems, suitably maybe adapted so as to include multiple reservoirs therein. Accordingly,the number of samples that suitably may be stored, retrieved, andanalyzed or used in acoustic ejection using conventional systems may bedramatically increased, and the efficiency of such storage, retrieval,analysis, and use also may be dramatically increased. Additionally, itshould be noted that other, non-acoustic liquid handling procedures alsosuitably may be used for extracting samples from the present reservoirs,such as with a pipette, capillary, pin tool, or other method, e.g.,based upon the insertion of a solid transfer device into a reservoir toextract a volume of the sample for transfer to another location.

Note that if an advantage of conventional tubes is the flexibility theyprovide in being selectable and separable from neighboring tubes, thenit may seem counterintuitive instead to configure a tube so as toinclude multiple reservoirs therein, that may be inseparable from oneanother. However, one desirable commercial use of fluids is the abilityto relatively quickly and conveniently construct dose-response curves,e.g., where the fluid contains a compound such as a drug. In anexemplary embodiment of the present invention, storing multipleconcentrations of a fluid within respective reservoirs within the sametube may speed a process of constructing the same assay but with of manydifferent concentrations of the fluid, e.g., compound or drug, withoutthe need for the separate retrieval of multiple, separable tubes.Additionally, because dose-response curves suitably may be constructedwith relatively small amounts of the same compound or drug usingacoustics, the consumption of any given concentration of the compoundmay be relatively small. In some embodiments, placing multipleconcentrations of the compound in the same tube may simplify retrieval,because retrieval and preparation of the tube suitably may supplymultiple concentrations, e.g., 4 concentrations. Additionally, such atube configuration may make better use of storage space as it providesome multiple of the concentrations, e.g., 4× the number ofconcentrations, in the same space relative to what may be achieved usingotherwise similar tubes having only a single reservoir therein.

Additionally, the present tubes optionally may be stored in collectionswith one another, e.g., stacked upon one another. For example, in someembodiments, the heights of the present tubes may be larger thanrequired to store the fluid therein, which may facilitate stacking. Forexample, the present tubes may be configured to as to receive caps, andthe top of the cap on one tube suitably may be configured so as toengage with the bottom of another tube, e.g., to interlock with thebottom of another tube. In some embodiments, the tube with which thepresent tube may be interlocked also may have multiple reservoirstherein, thus enabling further multiplication of the number of fluidsthat may be retrieved using a single act of retrieval. In oneillustrative embodiment, two stacked tubes each may include fourreservoirs defined therein, and each such reservoir may include adifferent concentration of a fluid. Accordingly, retrieving such a stackmay retrieve eight fluid concentrations at once, thus facilitating theready preparation of an 8-point dose response based on a singleretrieval. Note that any suitable number and types of tubes may be mixedand matched with one another. For example, a given stack may include anydesired number of tubes that include multiple reservoirs therein, e.g.,that include four reservoirs therein, may include any desired number oftubes that contain a different number of reservoirs therein, e.g., thatinclude two reservoirs therein, and may include any desired number oftubes that include only a single reservoir therein. For example, someexisting storage systems originally may have been designed toaccommodate an array of large tubes, but suitably may be used instead topermit storage of an array of stacks each of a desired number of thepresent tubes with caps, e.g., stacks of two or more, or three or more,or four or more, or five or more, or even ten or more of the presenttubes, wherein one or more of such tubes may include a plurality ofreservoirs, e.g., four reservoirs.

Note that any suitable number of reservoirs may be provided within thepresent sample tubes, and that any suitable one or more fluids may bestored within one or more of such reservoirs. For example, as notedabove, such fluids may have various concentrations of a compound, e.g.,a drug. The dilution sequence may include any desirable range ofconcentrations. For example, in some embodiments, the variousconcentrations may be provided in half-log steps. However, it should beappreciated that the concentrations may be provided in 10× steps, andmay be used for 16 point, half-log steps by using 1 drop as well as 3drops to fill in the intermediate half-log steps between the full logstep of concentration in the tubes.

Accordingly, the present tubes (containers) having multiple reservoirstherein suitably may facilitate the use of existing tube storageequipment, which originally may have been designed for use with largertubes, for storage and retrieval of the present tubes, and thus mayincrease storage efficiency and utility for dose response construction,among other potential applications.

FIGS. 1A-1B are simplified diagrams showing a single retrievable samplecontainer of roughly cylindrical shape having more than one reservoirwhere the samples are isolated from each other, optionally including anacoustically distinguishable difference in thickness in the bottoms ofreservoirs in the container, according to some embodiments of thepresent invention. These diagrams are merely examples, as are the otherdiagrams herein, which should not unduly limit the scope of the claims.One of ordinary skill in the art would recognize many variations,alternatives, and modifications.

FIG. 1A illustrates an exemplary container 100 that has a generallycylindrical shape, which also may be referred to herein as a “roughly”cylindrical shape, e.g., has a cross section that is generally circular.However, it should be appreciated that the cross-section of container100 suitably may have other geometries, e.g., may have a cross-sectionthat is generally triangular, or that is generally rectangular, or thatis generally square, or that is generally pentagonal, or that isgenerally hexagonal, or that has any suitable number of sides, andindeed may have any regular or irregular shape. Additionally, thecross-section of container 100 may vary along major axis 104. Forexample, container 100 may be tapered, e.g., such that the cross-sectionat one end of container 100 may be smaller or larger than thecross-section at the other end of container 100. For example, ifcontainer 100 is formed using a mold, then such a taper may facilitateremoval of container 100 from the mold.

A plurality of reservoirs 101 may be defined within container 100, e.g.,may extend along at least a portion of container 100 in a directionparallel to major axis 104. Preferably, reservoirs 101 may have has across section that is generally circular. However, it should beappreciated that the cross-section of reservoirs 101 suitably may haveother geometries, e.g., may have a cross-section that is generallytriangular, or that is generally rectangular, or that is generallysquare, or that is generally pentagonal, or that is generally hexagonal,or that has any suitable number of sides, and indeed may have anyregular or irregular shape. Additionally, the cross-section of one ormore of reservoirs 101 may vary along major axis 104. For example, oneor more of reservoirs 101 may be tapered, e.g., such that thecross-section at one end of reservoir 101 may be smaller or larger thanthe cross-section at the other end of that reservoir. For example, ifreservoirs 101 are formed using a mold, then such a taper may facilitateremoval of container 100 and reservoirs 101 from the mold. It should beunderstood that container 100 may include any suitable number ofreservoirs 101, e.g., two or more, or three or more, or four or more, orfive or more, or six or more, or ten or more, reservoirs 101.Additionally, it should be understood that each of such reservoirs 101within container 100 may have a shape and size that is selectedindependently of one or more other of such reservoirs. Preferably, eachof the reservoirs defined within the container 100 may be designed foracoustic ejection. Additionally, each of reservoirs 101 optionally maybe configured so as to be distinguishable acoustically from one another,so as to facilitate unique identification of the reservoirs withincontainer 100. For example, as illustrated in FIG. 1B, each ofreservoirs 101 optionally may be configured such that the bottom ofcontainer 101 has a different thickness beneath the respectivereservoir. For example, the bottom of container 100 may have a thicknessh1 beneath a first reservoir 101, and may have a thickness h2 beneath asecond reservoir 101. Such different thicknesses optionally may be usedso as to identify the individual reservoirs 101 within container 100,e.g., so as to identify the concentration of fluid within suchreservoirs 101, in a manner described in greater detail below withreference to FIG. 2.

Referring again to FIG. 1A, reservoirs 101 each respectively may definea volume configured to receive a fluid, e.g., a fluid into whichacoustic energy may be transmitted so as to eject a droplet of fluid. Inexemplary embodiments, container 100 has a height of 10 cm or less, or 6cm or less, or 2 cm or less, or 1 cm or less, or 1 mm or less, or 100 μmor less. In exemplary embodiments, container 100 has a diameter of 5 cmor less, or 2 cm or less, or 1 cm or less, or 0.5 cm or less, or 1 mm orless. In exemplary embodiments, each of reservoirs 101 independently mayhave a diameter of 1 cm or less, or 0.5 cm or less, or 1 mm or less, or100 μm or less.

In some embodiments, one or more of the reservoirs has a volume that isdifferent than a volume of at least one other of the reservoirs. Forexample, the reservoirs respectively may be configured to hold differentvolumes of fluid than one another. Illustratively, the difference involume can be significant, e.g., one of the reservoirs can have a volumethat is at least 10% larger than another of the reservoirs, e.g., canhave a volume that is at least 50% larger than another of thereservoirs, e.g., can have a volume that is at least 100% larger thananother of the reservoirs, e.g., can have a volume that is at least 400%larger than another of the reservoirs.

Preferably, the material used to provide container 100 is compatiblewith the fluid or fluids intended respectively to be contained withinreservoirs 101. Thus, if it is intended that the reservoirs 101 containan organic solvent such as acetonitrile, polymers that dissolve or swellin acetonitrile would be unsuitable for use in forming container 100.Similarly, reservoirs intended to contain dimethyl sulfoxide (DMSO)preferably are compatible with DMSO. For water-based fluids, a number ofmaterials are suitable for the construction of containers and include,but are not limited to, ceramics such as silicon oxide and aluminumoxide, metals such as stainless steel and platinum, and polymers such aspolyester and polytetrafluoroethylene. For fluids that arephotosensitive, the container 100 may be constructed from an opticallyopaque material that has sufficient acoustic transparency forsubstantially unimpaired functioning of the device. The container 100and reservoirs 101 therein may be prepared using any suitable technique,such as molding, machining, casting, extruding, or three-dimensionalprinting. As mentioned above, a cap suitably may be applied to thecontainer 100 at a later time so as to form a closed container thatencloses fluid.

FIG. 2 is a simplified diagram showing a retrievable sample container ofroughly cylindrical shape having more than one reservoir where thesamples are isolated from each other, in conjunction with an acousticejection system, according to some embodiments of the present invention.

In the embodiment illustrated in FIG. 2, container 200 is preferablyaxially symmetrical, e.g., cylindrical, having a plurality of sidewalls211 extending upward from container base 206, defining a plurality ofreservoirs 201 within container 200, and terminating at a plurality ofopenings 207, although other container shapes may be used. The materialand thickness of container base 206 preferably is configured such thatacoustic radiation may be transmitted therethrough and into the fluidsrespectively contained within reservoirs 101. Additionally, as notedabove with reference to FIG. 1B, the thickness of container base 206optionally may be different beneath each of reservoirs 201 so as tofacilitate identification of each of such reservoirs using the acousticejection system. For example, the time of flight of sound reflecting offof the bottom of reservoirs may be used to distinguish one reservoirfrom the other. For example, under a first reservoir the bottom 206 mayhave a thickness h1 which may be the designed to be the thinnest, undera second reservoir the bottom 206 may have a thickness h2 which isthicker than h1, and so on around counter-clockwise (or in any otherdesired arrangement) under any other reservoirs within container 200.When the container is presented to the acoustic ejector 240 such asdescribed in greater detail below, sonar measurements based on acousticreflections from the bottoms of reservoirs 201 may be used to determinewhich reservoir in the container is which and allow for the transducerto center itself relative to each of the reservoirs.

In this regard, it should be appreciated that an exemplary advantage ofacoustically readable identification and/or alignment marks, e.g.,different thicknesses of bottom 206 beneath each reservoir, is that suchmarks may for example be made compatible to the same transducer used forejection from the container and allow both sensing and actuation to beperformed by the same device or by similar device external to thetransfer station that would record or re-orient the container topre-determined configuration. However, alternative embodiments includeother alignment marks that may not necessarily be acoustically read andrather, are well known in the mechanical arts such as a key-way or useof optical marks (like barcodes or fiducials) to determine therotational orientation and identity of the wells in the container. Itshould be appreciated that many other patterns or acousticallydistinguishable features would be possible to suit the number of wells.

Preferably, but not necessarily, container base 206 is substantiallyflat, is oriented substantially perpendicular to the major axis ofsidewall 211, and is configured to receive an acoustic wave and totransmit the acoustic wave to fluid 203 disposed within a selected oneof reservoirs 201. For example, container 200 may be coupled to anacoustic ejector 240 that includes an acoustic radiation generator 241for generating acoustic radiation and an acoustic lens 243 for focusingthe acoustic radiation at a focal point within fluid 203 from which adroplet is to be ejected, near surface 204. The acoustic radiationgenerator contains a transducer 242, e.g., a piezoelectric element,commonly shared by an analyzer. In the illustrated embodiment, acombination unit 245 is provided that both serves as a controller and acomponent of an analyzer. For example, operating as a controller, thecombination unit 245 may provide the piezoelectric element 242 withelectrical energy that is converted into mechanical and acoustic energy.Or, for example, operating as a component of an analyzer, thecombination unit may receive and analyze electrical signals from thetransducer. The electrical signals may be produced as a result of theabsorption and conversion of mechanical and acoustic energy by thetransducer. Optionally, combination unit 245 also is configured so as toanalyze acoustic echoes reflected from the bottoms of reservoirs 201 soas to identify each of such reservoirs, e.g., based on thetime-of-flight of such acoustic echoes. Additionally, and independentlyof whether the thickness of bottom 206 varies beneath respectivereservoirs 201, combination unit 245 optionally may be configured so asto find the center of each of reservoirs 201 via sonar. For example,acoustic ejector 240 may include a motion system (not specificallyillustrated) configured to move acoustic radiation generator 241,acoustic lens 243, and acoustic coupling medium 244 relative tocontainer 200, or vice versa. In embodiments in which the location ofeach reservoir 201 was not otherwise made available to combination unit245, e.g., via pre-programming, combination unit 245 optionally may beconfigured so as to identify the center of each of reservoirs 201 usingacoustic echoes from the bottoms of reservoirs 201, and to control themotion system so as to move acoustic ejector 240 relative to container200 so as to center the point of focus of acoustic lens 243 at anappropriate point within a desired reservoir 201. Combination unit 245further may be configured so as to control the motion system so as tosubsequently move acoustic ejector 240 relative to container 200 so asto center the point of focus of acoustic lens 243 at an appropriatepoint within one or more different desired reservoirs 201, e.g., so asto eject droplets of fluid from each of reservoirs 201.

As shown in FIG. 2, acoustic lens 243 may include a single solid piecehaving a concave surface for focusing acoustic radiation, but the lensmay be constructed in other ways such as known in the art. Acousticejector 240 thus may be adapted to generate and focus acoustic radiationso as to eject a droplet of fluid from surface 204 when acousticallycoupled to container 200, and thus to fluid 203. Acoustic radiationgenerator 241 and lens 243 may function as a single unit controlled by asingle controller, or they may be independently controlled, depending onthe desired performance of the device. Typically, single ejector designsare preferred over multiple ejector designs because accuracy of dropletplacement and consistency in droplet size and velocity are more easilyachieved with a single ejector.

There are also a number of ways to acoustically couple the ejector 240to each individual reservoir 201 and thus to the fluid therein. One suchapproach is through direct contact as is described, for example, in U.S.Pat. No. 4,308,547 to Lovelady et al., wherein a lens constructed from ahemispherical crystal having segmented electrodes is submerged in afluid to be ejected. The aforementioned patent further discloses thatthe lens may be positioned at or below the surface of the fluid.However, this approach for acoustically coupling the lens to a fluid isundesirable when the ejector is used to eject different fluids in aplurality of containers or reservoirs, as repeated cleaning of the lenswould be required in order to avoid cross-contamination. The cleaningprocess would necessarily lengthen the transition time between eachdroplet ejection event. In addition, in such a method, fluid wouldadhere to the ejector as it is removed from each container, wastingmaterial that may be costly or rare.

Thus, one exemplary approach would be to acoustically couple the ejectorto the container without contacting any portion of the ejector, e.g.,lens 243, with any of the fluids to be ejected. To this end, ejector 240suitably may be positioned in controlled and repeatable acousticcoupling with container 200 to respectively eject droplets fromreservoirs 201 therein, without submerging the ejector therein.

For example, acoustic coupling may be achieved between the ejector andcontainer 200 through indirect contact, such as illustrated in FIG. 2.For example, acoustic coupling medium 244 may be placed between ejector240 and base 206 of container 200, with the ejector and containerlocated at a predetermined distance from each other. The acousticcoupling medium 244 may be an acoustic coupling fluid, preferably anacoustically homogeneous material in conformal contact with bothacoustic lens 243 and base 206 of container 200. Preferably, acousticcoupling medium 244 is substantially free of material having differentacoustic properties than the fluid medium itself. Furthermore, it ispreferred that acoustic coupling medium 244 includes a material havingacoustic properties, e.g., acoustic impedance, that facilitate thetransmission of acoustic radiation from acoustic lens 243 to bottomsurface 206 and into container 200 without significant attenuation inacoustic pressure and intensity. For example, as illustrated in FIG. 2,acoustic coupling medium 244 may couple container 200 to acoustic lens243, such that an acoustic wave generated by acoustic radiationgenerator 241 is directed by the lens 243 into the acoustic couplingmedium 244 which then transmits the acoustic wave into the container200. For example, combination unit 245 may control a motion system (notspecifically illustrated) so as to center lens 243 beneath a selectedone of reservoirs 201. The acoustic wave preferably focuses to a focalpoint 208 near the surface 204 of fluid 203 within that reservoir 201 inorder to eject at least one droplet 209 of the fluid. For furtherdetails of exemplary acoustic ejection systems and uses thereof, seeU.S. Pat. Nos. 6,938,995 and 7,900,505.

FIGS. 3A-3C illustrate exemplary configurations for storing samplecontainers with multiple reservoirs therein either singly or incombination with other containers, e.g., stacked with other containers,according to some embodiments of the present invention. For example,FIG. 3A illustrates an exemplary embodiment in which a sample containerthat includes multiple reservoirs therein, e.g., four reservoirs, may bestored separately from other containers. FIG. 3B illustrates anotherexemplary embodiment in which a pair of sample containers, each of whichindependently may include one or multiple reservoirs therein, e.g., fourreservoirs each, may be stored together in a collection, e.g., may bestacked. FIG. 3C illustrates yet another exemplary embodiment in whichthree sample containers, each of which independently may include one ormultiple reservoirs therein, e.g., four reservoirs in two of thecontainers, and one reservoir in another of the containers, may bestored together in a collection, e.g., may be stacked. As noted above,such collecting, e.g., stacking may facilitate efficient storage,retrieval, and analysis of fluids. For example, in the stack illustratedin FIG. 3B or the stack illustrated in FIG. 3C, the various reservoirswithin the stack optionally may include different concentrations of thesame fluid. Accordingly, retrieving one of such stacks suitably mayretrieve such concentrations, thus facilitating rapid analysis andacoustic ejection, e.g., may facilitate rapid preparation ofdose-response curves. Alternatively, one or more of the reservoirswithin the containers in a given stack may store fluids that areunrelated to one another, and as such, retrieving one of such stackssuitably may retrieve such unrelated fluids. As may be seen in FIGS.3B-3C, each container optionally may include a cap, and the cap of onecontainer may engage with, e.g., may interlock with, the base of anothercontainer.

FIG. 4A shows illustrates an exemplary rack for storing combinations,e.g., stacks of roughly cylindrical containers with multiple reservoirstherein for storage and retrieval, and FIG. 4B illustrates an exemplaryrack for individually holding roughly cylindrical containers withmultiple reservoirs therein for analysis or acoustic ejection, accordingto some embodiments of the present invention. These containers may bestored sealed or lidded (capped) with single-use seals or more preferredembodiment of a multi-use lid (cap) or multi-use septum which could beopened to facilitate acoustic transfer. Preferably, but not necessarily,the containers and lids are stackable with a roughly cylindricalexterior. Also, the de-stacking, de-lidding, re-lidding and re-stackingof the containers and lids may be performed in a variety of ways knownto those of skill in the art, and it is contemplated that other methodsmay emerge in the future that may also be used with the presentinvention to facilitate the retrieval, stacking, or destacking processesand preservation of materials within the container. In the exemplaryembodiment illustrated in FIG. 4A, a plurality of containers 400 havingmultiple reservoirs therein may be stacked and stored within each of aplurality of apertures 402 defined within a rack 401. At a desired time,one or more stacks of containers 400 may be removed and the containersthen may be disposed (de-stacking and de-lidding as needed) withinrespective spaces within rack 490 for acoustic analysis or ejection.Following such acoustic analysis or ejection, the containers then may bere-stacked and/or re-lidded as needed within an aperture defined withinrack 401 for storage until a later time. See, e.g., European PatentPublication No. EP 1348485, the entire contents of which areincorporated herein by reference for all purposes, for examples ofsuitable racks mechanisms for manipulating stacks of containers.

FIG. 5A-5B are simplified diagrams showing exemplary collections ofmultiple sample containers surface with their corresponding seals (orlids or caps) stacked as a linked assembly with roughly cylindricalexterior for storage or retrieval with different concentrations in eachwell, according to some embodiments of the present invention. In theembodiment illustrated in FIG. 5A, 3 sample containers are shown in astacked configuration wherein each container has a lid and the compositeexterior for the containers is roughly cylindrical. The retrieval of astack of containers as shown in the specific example of FIG. 5A shows amixed stack of containers with one containing a high-concentration of acompound in solution in the top, single-reservoir container designated1×, and lower concentrations of the compound in solution in each of themulti-reservoir containers wherein the concentration is designated by3×, 10× and so on to reflect the level of dilution. This roughlycorresponds to half-log dilution steps for the concentrations. Withthese containers, the span of concentrations is 4 logs or 1× to 10,000×.It should be appreciated that these concentrations, or any other desiredconcentrations, suitably may be arranged in any manner within thevarious wells of the stack of containers. For example, differentconfigurations are shown in FIGS. 4A and 4B where one might be moreamenable for transferring in a pre-made serial dilution (e.g., FIG. 4A)in and the other for dilution construction from one container to theother (e.g., FIG. 4B). For example, in one illustrative embodiment, 1part of a well in one container could be mixed with 2 parts diluent in awell in the other container to construct a 1:3 step between the twolower containers of the stack such as in the illustrative embodimentshown in FIG. 5B. Retrieving a container stack with a serial dilutionseries such as in FIG. 5B may facilitate rapid construction of a 4-logspan and half-log step assay, for example, when equal volumes of each ofthese was is transferred to corresponding assay wells. While illustratedwith a half log step, the step in storage concentrations suitably may bea different number, larger or smaller, such as 2 or 10. In the case of10, or full log steps, intermediate concentrations can be constructed byvarying the amount transferred from the wells to fill in the half logsteps. For example, if the containers hold in any order 1×, 10×, 100×,and so on, corresponding to log steps in concentrations, then doseresponse assay with half-log steps may be constructed in a series ofassay wells through the acoustic transfer of a different number ofdroplets. In such an example, a droplet from the 1× concentration wellmay form the highest concentration assay well, 3 droplets of the samevolume as the 1× droplet but transferred from the 10× dilution may formthe 2^(nd) highest concentration assay well, 1 droplet of the samevolume as the previous one from the 10× dilution to the 3^(rd) highestconcentration destination assay well, 3 droplets from the 100× dilutionmay form the next assay well, and so on. It should be appreciated thatthose of skill in the art could design many configurations of differentconcentrations or dilutions (e.g., in even log steps such as describedabove, or other dilutions), whose intermediate values were filled inwith various volumes to construct dose response assays. It is alsocontemplated that the reservoirs could also be used to carry the diluentor control samples or other materials required for the construction ofthe assay. In one example, the samples within a given container, orwithin a collection, e.g., stack of containers, may be related to oneanother. For example, the relationship may be from fractionation (e.g.,centrifugation), liquid chromatography (LC) effluent, a time-series ofsamples obtained from a patient, or DNA samples from close relatives,among others.

Note that in some embodiments, it may be desirable to rotate the presentcontainers so as to arrange the reservoirs therein in the proper placefor acoustic analysis or ejection. Suitable methods for sucharrangements, e.g., geometric features in the container construction toreadable visual methods, suitably may be adapted for use with thepresent containers. Additionally, as noted above, one way to allowidentification of wells is to have acoustically distinguishable featuresin the tube. For example, such features may be present in the bottom ofthe container and designed to be detected when the tube was presented toan acoustic reader or a transfer device. One preferred embodiment wouldbe to have one or more of the wells at measurably different thicknessfrom the others to provide a reference for which well is which in thetube. The time of flight for sound in between the bottom of the tube andbottom of each reservoir could then be used to determine theorientation/identity of the reservoir within the tube (e.g., is itreservoir #1, #2, #3 or #4). Each of these separable multi-reservoirtubes could also be provided with an identification mark such asdescribed in provisional patent application No. 61/927,395, filed Jan.14, 2014 and entitled “Sample Containers Having Identification MarksEmbedded Therein and Being Adapted for Acoustic Ejections,” the entirecontents of which are incorporated by reference herein for all purposes.For example, FIG. 4B illustrates an exemplary identification mark 480disposed on each of containers 400. It should be understood that use ofsuch identification marks is purely optional.

Under one non-limiting aspect of the present invention, a generallytubular container includes a plurality of reservoirs defined therein.Illustratively, such a container is described above with reference toFIGS. 1A, 1B, 2, 3A, 3B, 3C, 4A, 4B, 5A, and 5B.

In some embodiments, the generally tubular container is adapted foracoustic ejection of a fluid disposed within at least one of thereservoirs of the plurality of reservoirs.

In some embodiments, a plurality of fluids are respectively disposedwithin the reservoirs of the plurality of reservoirs. Optionally, theplurality of fluids are selected from the group consisting of: differentsamples from the same patient, different fractions of a sample from apatient, and different concentrations of the same compound, chemical, ordrug.

In some embodiments, the reservoirs isolate the plurality of fluids fromone another.

In some embodiments, each reservoir includes a bottom, the bottom of atleast one of the reservoirs including an acoustically distinguishabledifference in thickness from the bottom of at least one other of thereservoirs.

In some embodiments, the generally tubular container is adapted forextraction of a fluid disposed within at least one of the reservoirs ofthe plurality of reservoirs using a non-acoustic liquid handling method.In some embodiments, the non-acoustic liquid handling method is based onthe insertion of a solid transfer device into one of the reservoirs. Insome embodiments, the solid transfer device includes a pipette,capillary, or pin tool.

In some embodiments, the plurality of reservoirs include three or morereservoirs.

In some embodiments, a cross-section of the generally tubular containeris generally circular.

In some embodiments, one or more of the reservoirs has a volume that isdifferent than a volume of at least one other of the reservoirs. Forexample, the reservoirs respectively may be configured to hold differentvolumes of fluid than one another. Illustratively, the difference involume can be significant, e.g., one of the reservoirs can have a volumethat is at least 10% larger than another of the reservoirs, e.g., canhave a volume that is at least 50% larger than another of thereservoirs, e.g., can have a volume that is at least 100% larger thananother of the reservoirs, e.g., can have a volume that is at least 400%larger than another of the reservoirs.

In some embodiments, the generally tubular container includes a majoraxis, each of the reservoirs extending along at least a portion of thecontainer in a direction parallel to the major axis.

In some embodiments, the container is tapered along the major axis.

Under another non-limiting aspect of the present invention, a stackedplurality of generally tubular containers is provided, at least one ofthe generally tubular containers including a plurality of reservoirsdefined therein. Illustratively, such a stacked plurality of generallytubular containers is described above with reference to FIGS. 3B, 3C,4A, 5A, and 5B. Illustratively, such a container is described above withreference to FIGS. 1A, 1B, 2, 3A, 3B, 3C, 4A, 4B, 5A, and 5B.

In some embodiments, the stacked plurality of generally tubularcontainers is retrievable as a group.

In some embodiments, the at least one of the generally tubularcontainers is adapted for acoustic ejection of a plurality of fluidsrespectively disposed within the reservoirs of the plurality ofreservoirs.

Under another non-limiting aspect of the present invention, a methodincludes extracting a fluid from a generally tubular container includinga plurality of reservoirs defined therein. Illustratively, such a methodis described above with reference to FIG. 2. Illustratively, such acontainer is described above with reference to FIGS. 1A, 1B, 2, 3A, 3B,3C, 4A, 4B, 5A, and 5B.

In some embodiments, the extracting comprises acoustically ejecting thefluid.

In some embodiments, the fluid is disposed within at least one of thereservoirs of the plurality of reservoirs.

In some embodiments, a plurality of fluids are respectively disposedwithin the reservoirs of the plurality of reservoirs. Optionally, theplurality of fluids are selected from the group consisting of: differentsamples from the same patient, different fractions of a sample from apatient, and different concentrations of the same compound, chemical, ordrug.

In some embodiments, extracting a fluid disposed within at least one ofthe reservoirs of the plurality of reservoirs comprises using anon-acoustic liquid handling method. In some embodiments, thenon-acoustic liquid handling method is based on the insertion of a solidtransfer device into one of the reservoirs. In some embodiments, thesolid transfer device includes a pipette, capillary, or pin tool.

In some embodiments, one or more of the reservoirs has a volume that isdifferent than a volume of at least one other of the reservoirs. Forexample, the reservoirs respectively may be configured to hold differentvolumes of fluid than one another. Illustratively, the difference involume can be significant, e.g., one of the reservoirs can have a volumethat is at least 10% larger than another of the reservoirs, e.g., canhave a volume that is at least 50% larger than another of thereservoirs, e.g., can have a volume that is at least 100% larger thananother of the reservoirs, e.g., can have a volume that is at least 400%larger than another of the reservoirs.

In some embodiments, each reservoir includes a bottom, the bottom of atleast one of the reservoirs including an acoustically distinguishabledifference in thickness from the bottom of at least one other of thereservoirs.

Some embodiments further include acoustically distinguishing the atleast one of the reservoirs from the at least one other of thereservoirs.

In some embodiments, the reservoirs isolate the plurality of fluids fromone another.

In some embodiments, the plurality of reservoirs including three or morereservoirs.

In some embodiments, a cross-section of the generally tubular containeris generally circular.

In some embodiments, the generally tubular container includes a majoraxis, each of the reservoirs extending along at least a portion of thecontainer in a direction parallel to the major axis.

In some embodiments, the container is tapered along the major axis.

Under another non-limiting aspect of the present invention, a methodincludes retrieving, as a group, a stacked plurality of generallytubular containers, at least one of the generally tubular containersincluding a plurality of reservoirs defined therein. Illustratively,such a method is described above with reference to FIGS. 3B, 3C, 4A, 5A,and 5B. Illustratively, such a container is described above withreference to FIGS. 1A, 1B, 2, 3A, 3B, 3C, 4A, 4B, 5A, and 5B.

In some embodiments, the at least one of the generally tubularcontainers is adapted for acoustic ejection of at least one fluiddisposed within at least one of the reservoirs of the plurality ofreservoirs.

All patents, patent applications, and publications mentioned herein arehereby incorporated by reference in their entireties for all purposes.However, where a patent, patent application, or publication containingone or more express definitions is incorporated by reference, thoseexpress definitions should be understood to apply to the incorporatedpatent, patent application, or publication in which the one or moreexpress definitions are found, but not to the remainder of the text ofthis application, in particular not to the claims of this application.

Although specific embodiments of the present invention have beendescribed, it will be understood by those of skill in the art that thereare other embodiments that are equivalent to the described embodiments.Accordingly, it is to be understood that the invention is not to belimited by the specific illustrated embodiments, but only by the scopeof the appended claims.

1.-35. (canceled)
 36. A method for extracting fluid from a plurality ofreservoirs of a container, the method comprising: positioning anacoustic ejector relative to a first reservoir of the plurality ofreservoirs such that a first acoustic beam of the acoustic ejector isdirected toward the first reservoir; ejecting a first droplet from thefirst reservoir; re-positioning the acoustic ejector relative to asecond reservoir of the plurality of reservoirs such that a secondacoustic beam of the acoustic ejector is directed toward the secondreservoir; and ejecting a second droplet from the second reservoir. 37.The method of claim 36, wherein the first reservoir comprises a compoundat a first concentration and wherein the second reservoir comprises thecompound at a second concentration, and wherein the method furthercomprises constructing a dose-response curve using at least the firstdroplet and the second droplet.
 38. The method of claim 36, wherein thefirst reservoir comprises a first compound, wherein the second reservoircomprises a second compound; and wherein the first compound is differentthan the second compound.
 39. The method of claim 36, further comprisinga motion system configured to: move the acoustic ejector to position theacoustic ejector relative to the first reservoir such that the firstacoustic beam is directed toward the first reservoir; and move theacoustic ejector to re-position the acoustic ejector relative to thesecond reservoir such that the second acoustic beam is directed towardthe second reservoir.
 40. The method of claim 39, further comprising acontroller, wherein a first location of the first reservoir is receivedby the controller and sent to the motion system prior to the motionsystem moving the acoustic ejector to position the acoustic ejectorrelative to the first reservoir; and wherein a second location of thesecond reservoir is received by the controller and sent to the motionsystem prior to the motion system moving the acoustic ejector tore-position the acoustic ejector relative to the second reservoir. 41.The method of claim 39, further comprising a controller, wherein thecontroller is configured to identify a first center of the firstreservoir using at least one first acoustic echo from a first bottom ofthe first reservoir in order to move the acoustic ejector to positionthe acoustic ejector relative to the first reservoir; and wherein thecontroller is configured to identify a second center of the secondreservoir using at least one second acoustic echo from a second bottomof the second reservoir in order to move the acoustic ejector tore-position the acoustic ejector relative to the second reservoir. 42.The method of claim 36, further comprising a motion system configuredto: move the container to position the first reservoir relative to theacoustic ejector such that the first acoustic beam is directed towardthe first reservoir; and move the container to re-position the secondreservoir relative to the acoustic ejector such that the second acousticbeam is directed toward the second reservoir.
 43. The method of claim42, further comprising a controller, wherein a first location of thefirst reservoir is received by the controller and sent to the motionsystem prior to the motion system moving the container to position theacoustic ejector relative to the first reservoir; and wherein a secondlocation of the second reservoir is received by the controller and sentto the motion system prior to the motion system moving the container tore-position the acoustic ejector relative to the second reservoir. 44.The method of claim 42, further comprising a controller, wherein thecontroller is configured to identify a first center of the firstreservoir using at least one first acoustic echo from a first bottom ofthe first reservoir in order to move the container to position theacoustic ejector relative to the first reservoir; and wherein thecontroller is configured to identify a second center of the secondreservoir using at least one second acoustic echo from a second bottomof the second reservoir in order to move the container to re-positionthe acoustic ejector relative to the second reservoir.
 45. The method ofclaim 36, further comprising a motion system, wherein the container isstacked with at least one other container, and wherein the motion systemis configured to de-stack the container from the at least one othercontainer prior to positioning the acoustic ejector relative to thefirst reservoir.
 46. The method of claim 45, wherein each reservoircomprises a lid, and wherein the motion system is configured to: de-lidthe first reservoir prior to ejecting the first droplet from the firstreservoir; and de-lid the second reservoir prior to ejecting the seconddroplet from the second reservoir.
 47. The method of claim 36, whereinthe container is a generally tubular container and wherein the containercomprises more than two reservoirs.
 48. The method of claim 47, whereineach reservoir of the plurality of reservoirs is cylindrical.
 49. Themethod of claim 36, wherein each reservoir of the plurality ofreservoirs comprises a different concentration of a compound.
 50. Amethod for extracting fluid from a plurality of reservoirs of acontainer, the method comprising: ejecting a first droplet from a firstreservoir of the plurality of reservoirs; ejecting a second droplet froma second reservoir of the plurality of reservoirs; and constructing adose-response curve using at least the first droplet and the seconddroplet.
 51. The method of claim 50, wherein the first droplet comprisesa first concentration of the compound, wherein the second dropletcomprises a second concentration of the compound, and wherein the firstconcentration is different than the second concentration.
 52. The methodof claim 50, wherein the first reservoir comprises a first compound,wherein a third reservoir comprises a second compound, and wherein thefirst compound is different than the second compound.
 53. The method ofclaim 50, wherein the container is a generally tubular container.
 54. Acontainer comprising a plurality of reservoirs, the containercomprising: a first reservoir of the plurality of reservoirs, whereinthe first reservoir comprises a first compound; and a second reservoirof the plurality of reservoirs, wherein the second reservoir comprises asecond compound.
 55. The container of claim 54, wherein the firstreservoir comprises a first concentration of the first compound and thecontainer further comprising a third reservoir of the plurality ofreservoirs, wherein the third reservoir comprises a second concentrationof the first compound.
 56. The container of claim 54, wherein thecontainer is a generally tubular container, and wherein each reservoirof the plurality of reservoirs is cylindrical.